Daphnane diterpenoid resistant to prostate cancer and preparation method thereof

ABSTRACT

The invention relates to a daphnane diterpenoid resistant to a prostate cancer and a preparation method thereof. A daphnane diterpenoid compound as represented by formula (I) or formula (II) significantly inhibits the proliferation of various prostate cancer cells; the activities of some such compounds at a cellular level and an animal level are higher than that of the existing clinical targeting drug enzalutamide; and the daphnane diterpenoid compounds have strong synergistic effects when used in combination with the enzalutamide and are expected to become candidate drugs for treatment or adjuvant treatment of a castration-resistant prostate cancer.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application is a Continuation in Part Application ofInternational Application PCT/CN2021/116955, filed Sep. 7, 2021, whichclaims the benefit of and priority to Chinese Patent Application No.202011524327.0, filed Dec. 22, 2020, the entire disclosures of which areincorporated herein by reference.

TECHNICAL FIELD

The present disclosure relates to a natural compound, a preparationmethod and use thereof, and in particular to a daphnane diterpenoidcompound, a preparation method and use thereof in preparing a medicamentresistant to castration-resistant prostate cancer.

BACKGROUND

Prostate cancer is a highly prevalent malignancy in middle-aged andelderly people. According to the latest cancer statistics from theAmerican Cancer Society (ACS) in 2020, the incidence rate of theprostate cancer in the United States ranks highest and the mortalityrate takes second among malignant tumors. In China, the incidence rateof prostate cancer is also soaring year by year with the changes in thestandard of living and the pace of life.

In the later stage, the prostate cancer evolves intocastration-resistant prostate cancer which is ineffective hormonetherapy, resulting in metastasis, which is an important factor leadingto patient death. The castration-resistant prostate cancer is closelyrelated to the abnormally activated androgen receptor (AR) signalingpathway. The anti-androgen drugs enzalutamide and abiraterone, whichwere marketed in 2010 and 2011 respectively, can significantly improvepatient survival. However, due to drug resistance, little effectivenessof treatment was achieved in the later stage. Therefore, the developmentof a new drug for the castration-resistant prostate cancer has importantresearch significance and application value.

Natural products, due to their structural diversity and goodbio-compatibility, are an important source for the research anddevelopment of the new drug. Literature research has shown that daphnanediterpenoids with a novel structure are abundant in Daphne plants. Thedaphnane diterpenoids are complex molecular structures with a 5/7/6tricyclic carbon skeleton, and comprises multiple chiral centers thatusually contain chiral hydroxyl groups at the positions of C3, C4, C5,C9, C13, C14, C20, etc., among which many compounds form a specificorthoester structure among the chiral hydroxyl groups at the C9, C13,and C14 positions.

Technical Problem

The daphnane diterpenoids have anti-HIV, anti-leukemia, anti-tumor,neuroprotective, insecticidal and cytotoxic activities, etc. Also, aresearch has found that such diterpenoids have a very significantinhibitory activity on prostate cancer cells, which is stronger than theexisting clinical targeting drug enzalutamide at a cellular level and ananimal level. However, the research on in-depth pharmacodynamics,structure-activity relationship and action mechanism of suchditerpenoids in resisting to the prostate cancer is still lacking.

SUMMARY

According to the first aspect of the present disclosure, there isprovided:

use of a daphnane diterpenoid compound in preparing a medicament fortreatment or combination treatment of a castration-resistant prostatecancer, wherein the daphnane diterpenoid compound has a general formalas represented by formula I or formula II:

in the formula I and the formula II, a bond between C-1 and C-2 is asingle bond or is a double bond, a bond between C-6 and C-7 is a singlebond or is a double bond, and a bond between C-15 and C-16 is a singlebond or is a double bond;

R₁ is selected from hydrogen or hydroxyl or R₁ is absent when a doublebond is formed between C-1 and C-2;

R₂ is selected from hydrogen, hydroxyl, carbonyl, benzoyl or acetyl;

R₃ is selected from hydrogen, hydroxyl, acetyl, isovaleryl, crotonyl orbenzoyl;

R₄ is selected from hydrogen, hydroxyl, acetyl, isobutyryl,2-thienylcarbonyl, benzoyl or palmitoyl;

R₅ is selected from hydroxyl, fluorine, chlorine, bromine, and iodine,and forms a ternary epoxy with R₆ or R₆ is absent when a double bond isformed between C-6 and C-7;

R₆ is selected from hydrogen, hydroxyl, fluorine, chlorine, bromine andiodine, and forms a ternary epoxy with R₅ or R₅ is absent when a doublebond is formed between C-6 and C-7;

in the formula I: R₇ is selected from methyl, phenyl, nonanyl, (1E,3E)-nonadienyl, (1E, 3Z)-nonadienyl or (1E, 3E, 5E)-nonatrienyl;

in the formula II: R₇ is selected from hydrogen, benzoyl, acetyl,decanoyl, (2E, 4E)-decadienoyl, (2E, 4Z)-decadienoyl or (2E, 4E,6E)-decatrienoyl;

R₈ is selected from hydrogen or hydroxyl or R₈ is absent when a doublebond is formed between C-15 and C-16; and

R₉ is selected from hydrogen, hydroxyl, acetyl, benzoyl, isobutyryl,butyryl or propionyl.

In some embodiments, the daphnane diterpenoid compound is selected from:

a compound of formula (III):

wherein the compound of formula (III) is selected from the groupconsisting of:

Compound R₁ R₂ R₃ R₄ R₇ R₈ R₉ YH-1 Δ1 ═O OH OH Ph Δ16 OBu YH-2 Δ1 ═O OHOH Ph Δ16 OiBu YH-3 Δ1 ═O OH OH Ph Δ16 OProp YH-4 Δ1 ═O OH OH Ph Δ16 OHYH-5 Δ1 ═O OH OG Ph Δ16 OAc YH-6 Δ1 ═O OH OH Ph Δ16 OAc YH-7 Δ1 ═O OAcOAc Ph Δ16 OAc YH-8 Δ1 ═O OH OAc Ph Δ16 OAc YH-9 Δ1 ═O OH OBz Ph Δ16 OAcYH-10 Δ1 ═O OH OS Ph Δ16 OAc YH-11 Δ1 ═O OH OH Ph Δ16 OBz YH-12 Δ1 ═OOAc OAc Ph Δ16 OBz YH-13 Δ1 ═O OH OAc Ph Δ16 OBz YH-14 Δ1 ═O OH OH A Δ16OH YH-15 Δ1 ═O OH OH A Δ16 OBu YH-16 Δ1 ═O OH OH A Δ16 OBz YH-17 Δ1 ═OOH OH D Δ16 OBz YH-18 Δ1 ═O OAc OAc A Δ16 OBz YH-19 Δ1 ═O OH OAc A Δ16OBz YH-20 Δ1 ═O OH OBz A Δ16 OBz YH-21 Δ1 ═O OH OS A Δ16 OAc YH-22 Δ1 ═OOH OH A Δ16 OAc YH-23 Δ1 ═O OAc OAc A Δ16 OAc YH-24 Δ1 ═O OH OAc A Δ16OAc YH-25 Δ1 ═O OH OBz A Δ16 OAc YH-26 Δ1 ═O OH OS A Δ16 OAc YH-27 Δ1 ═OOH OG A Δ16 OAc YH-28 Δ1 ═O OH OG D Δ16 OBz YH-29 Δ1 ═O OH OH B Δ16 OBzYH-30 Δ1 ═O OH OH Me Δ16 OAc YH-31 Δ1 ═O OH OH B Δ16 OAc YH-32 Δ1 ═O OHOH D Δ16 OAc YH-33 Δ1 ═O OH OH F Δ16 OAc YH-34 Δ1 ═O OH OH F Δ16 OBzYH-35 αH ═O OH OH Ph Δ16 OAc YH-36 αH ═O OH OH Ph Δ16 OBz YH-37 Δ1 ═O OHOH Da H OAc YH-38 αH ═O OH OH Da H OAc YH-39 αH ═O OH OH Ph H OBz YH-40Δ1 ═O OH OH Ph H OBz YH-41 Δ1 ═O OH OH Ph Δ16 H YH-42 αH βO(CO)E OH OHPh Δ16 H YH-43 αH βOH OH βOE Ph Δ16 H;a compound of formula (IV):

wherein the compound of formula (IV) is selected from the groupconsisting of:

Compound R₃ R₄ R₅ R₆ R₇ R₉ YH-44 H OH Δ6 H Me OBz YH-45 OH OBz βCl αOHPh OAc YH-46 OH OBz αOH βCl Ph OAc YH-47 OH OH βCl αOH Ph OAc YH-48 OHOH αOH βCl Ph OAc YH-49 OH OH βCl αOH Ph OBz YH-50 OH OH αOH βCl Ph OBzYH-51 OH OH αOH αOH Ph OBz YH-52 OH OH βBr αOH Ph OBz YH-53 OH OH αOHβBr Ph OBz;a compound of formula (V):

wherein the compound of formula (V) is selected from the groupconsisting of:

Compound R₇ R₉ YH-54 COA OAc YH-55 COA OBz YH-56 Bz OBz;anda compound of formula (VI):

wherein the compound of formula (VI) is selected from the groupconsisting of:

Compound R₇ R₉ YH-57 H OBz YH-58 H OAc YH-59 COD OAc YH-60 Bz OBz YH-61COA OH YH-62 COA OAc YH-63 COA OBz YH-64 COA H;andwherein in formula (III), formula (IV), formula (V), and formula (VI):

Δ1 is when R₁ is absent and a double bond is formed between C-1 and C-2;Δ6 is when R₅ is absent and a double bond is formed between C-6 and C-7;Δ16 is when R₈ is absent and a double bond is formed between C-15 andC-16;α is when the identified substituent is on the opposite face of thehydroxyl group of the contiguous ring; andβ is when the identified substituent is on the same face as the hydroxylgroup of the contiguous ring.

In some embodiments, the daphnane diterpenoid compound is at least oneselected from the group consisting of YH-6, YH-11, YH-16, YH-17, YH-22,YH-35, YH-36, YH-47, YH-48, YH-49, YH-50, YH-52 and YH-53.

In some embodiments, the daphnane diterpenoid compound further comprisesa pharmaceutically acceptable derivative thereof.

In some embodiments, the pharmaceutically acceptable derivative is asalt thereof.

According to the second aspect of the present disclosure, there isprovided:

a daphnane diterpenoid compound and a medicinal derivative thereof, thedaphnane diterpenoid compound is as defined in the first aspect of thepresent disclosure.

In some embodiments, the daphnane diterpenoid compound is selected fromthe compounds of YH-8, YH-9, YH-10, YH-19, YH-20, YH-21, YH-24, YH-25,YH-26, YH-30, YH-33, YH-34, YH-37, YH-38, YH-39, YH-45, YH-46, YH-47,YH-48, YH-49, YH-50, YH-52, YH-53, YH-56, YH-57, YH-60 and YH-61.

According to the third aspect of the present disclosure, there isprovided:

a composition for treatment or adjuvant treatment of acastration-resistant prostate cancer, an active ingredient of thecomposition comprises at least one of the daphnane diterpenoid compounddefined in the first aspect of the present disclosure and the acceptablymedicinal derivative thereof. In some embodiments, the pharmaceuticallyacceptable derivative is a salt thereof.

In some embodiments, the daphnane diterpenoid compound is selected fromthe compounds of YH-6, YH-11, YH-16, YH-17, YH-22, YH-35, YH-36, YH-8,YH-9, YH-10, YH-19, YH-20, YH-21, YH-24, YH-25, YH-26, YH-30, YH-33,YH-34, YH-37, YH-38, YH-39, YH-45, YH-46, YH-47, YH-48, YH-49, YH-50,YH-52, YH-53, YH-56, YH-57, YH-60 and YH-61, and an acceptably medicinalderivative thereof.

In some embodiments, the composition further comprises at least onecompound having a therapeutic effect on the prostate cancer, preferably,the compound having the therapeutic effect on the prostate cancer is atleast one selected from the group consisting of enzalutamide,abiraterone, cyclophosphamide, adriamycin, docetaxel and mitoxantrone.

According to the forth aspect of the present disclosure, there isprovided:

a method for treatment or adjuvant treatment of a castration-resistantprostate cancer, comprising:

detecting to confirm that a patient suffers from thecastration-resistant prostate cancer; and

administering to the patient a therapeutic amount of a daphnanediterpenoid compound or a pharmaceutically acceptable salt, solvate,co-crystal thereof, wherein the daphnane diterpenoid compound is asdefined in the first aspect of the present disclosure.

In some embodiments, the method further comprises administering to apatient at least one compound having a therapeutic effect on theprostate cancer, preferably, the compound having the therapeutic effecton the prostate cancer is at least one selected from the groupconsisting of enzalutamide, abiraterone, cyclophosphamide, adriamycin,docetaxel and mitoxantrone.

The present disclosure has the beneficial effects that: in the presentdisclosure, a daphnane diterpenoid compound is isolated or synthesisedfor the first time. Such compounds can significantly inhibit theproliferation of various prostate cancer cells; and in particular,Compound YH-52 has a higher activity at a cellular level and an animallevel than that of the existing clinical targeting drug enzalutamide. Inaddition, the combination of the daphnane diterpenoid compound withenzalutamide can obtain a strong synergistic effect, therefore suchcompounds are expected to become candidate drugs for treatment of acastration-resistant prostate cancer.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a flowchart showing the extraction and isolation of daphnegenkwa.

FIG. 2A shows the effect of oral administration of Compound YH-52 on thesize of tumor.

FIG. 2B shows the effect of oral administration of Compound YH-52 on thevolume of tumor.

FIG. 2C shows the effect of oral administration of Compound YH-52 on theweight of tumor.

FIG. 3A shows the inhibitory effect of intraperitoneal injectionadministration of Compound YH-52 combined with enzalutamide on prostatecancer cells 22RV1.

FIG. 3B shows the effect of intraperitoneal injection administration ofCompound YH-52 combined with enzalutamide on the size of tumor.

FIG. 3C shows the effect of intraperitoneal injection administration ofCompound YH-52 combined with enzalutamide on the volume of tumor.

FIG. 3D shows the effect of intraperitoneal injection administration ofCompound YH-52 combined with enzalutamide on the weight of tumor.

DETAILED DESCRIPTION

Thirty-four (34) natural daphnane diterpenoids were isolated by theinventors from thymelaeaceae plants, daphne genkwa, and 30 derivativeswere obtained from some compounds after being carried out with astructural modification. Relevant tests on the obtained series ofditerpenoids for resisting to prostate cancer cells found a series ofcompounds that have a significant inhibitory activity on tumor cells anda stronger effect than that of the existing clinical targeting drugenzalutamide, among which one of the new compounds is expected to be acandidate drug for the treatment of the castration-resistant prostatecancer.

The technical scheme of the present disclosure will be explained in moredetail with reference to the following embodiments. Unless otherwisespecified, the reagents, equipment, and methods used in the presentdisclosure are those routinely commercially available and routinely usedin the art.

In the present disclosure, research was carried out on thymelaeaceaeplants (taking daphne genkwa as an example).

Equipment and reagents: a Bruker AM-400/500 spectrometer was used forrecording NMR spectrum, and a TMS was used as an internal standard.Column chromatography silica gel (300-400 mesh): Qingdao HaiyangChemical Co., Ltd.; GF₂₅₄ silica gel thin-layer chromatographyprefabricated plate: Qingdao Haiyang Chemical Co., Ltd.; MCI filler(CHP20P, 75-150 μm): Mitsubishi Chemical Industries Ltd.; Sephadex(Sephadex LH-20): GE Company in USA; ODS filler (12 nm, S-50 μm): YMC.Co., Ltd in Japan; and other solvents and reagents: analytical pure(AR), Baishi Chemical Industry Co., Ltd, Tianjin, China.

Preparation of Daphnane Diterpenoid Compound

Twenty kg of daphne genkwa was taken, and crushed into coarse powder.Three times the volume of 95% ethanol was added, soaked for 1 week, andrecovered the ethanol under reduced pressure after suction filtration.The soaking and extraction steps were repeated more than three times,and finally 1500 g of the ethanol extract of daphne genkwa was obtained.The extract of daphne genkwa was dispersed with 1 L of water, and thenextracted three times with ethyl acetate. The ethyl acetate extractliquor were combined and concentrated under reduced pressure to obtain300 g of ethyl acetate extract.

The above ethyl acetate extract was taken and loaded on a silica gelcolumn, initially segmented with the mixture of petroleum ether andethyl acetate, the mixture of dichloromethane and methanol, then furtherisolated by ODS, MCI, Sephadex LH-20, finally purified by thesemi-preparative high-performance liquid chromatography under theconditions of (acetonitrile:water) or (methanol:water), and 34 monomercompounds were obtained. FIG. 1 shows the specific isolation flowchart.The obtained partial daphnane diterpenoid compounds were structurallymodified to prepare derivatives thereof

Identification of Isolated Products Example 1

Compound YH-30 was obtained by purifying with the semi-preparative HPLC.The structure and data of Compound YH-30 were as follows:

YH-30: [α]²⁵ _(D)+7.8 (c 0.230, CH₂Cl₂); UV (MeOH) Amax (log ε) 232(4.21) nm; ECD (c 3.3×10⁻⁴ M, MeCN) λ_(max) (Δε) 252 (−2.44) nm; IR(KBr) v_(max) 3445, 2925, 2856, 1705, 1632, 1451, 1401,1379, 1301, 1268,1108, 1070,1026, 938, 913, 863 cm⁻¹; ¹H NMR (CDCl₃, 400 MHz) δ_(H) 7.55(1H, s, H-1), 4.20 (1H, s, H-5), 3.61 (1H, s, H-7), 3.59 (1H, d, J=2.4Hz, H-8), 3.76 (1H, m, H-10), 2.52 (1H, q, J=7.2 Hz, H-11), 5.18 (1H, s,H-12), 4.84 (1H, d, J=2.4 Hz, H-14), 4.99 (2H, s, H-16), 1.84 (3H, s,H₃-17), 1.37 (3H, d, J=7.2 Hz, H₃-18), 1.76 (3H, br s, H₃-19), 3.81 (1H,d, J=12.0 Hz, H-20a), 3.90 (1H, d, J=12.0 Hz, H-20b), 1′-Me: 1.70 (3H,s); 12-OBz: 7.87 (2H, m), 7.44 (2H, m), 7.57 (1H, m); ¹³C NMR (CDCl₃,100 MHz) δ_(C) 160.3 (C-1), 136.9 (C-2), 209.4 (C-3), 72.2 (C-4), 71.8(C-5), 60.7 (C-6), 64.1 (C-7), 35.5 (C-8), 77.8 (C-9), 47.4 (C-10), 44.0(C-11), 78.9 (C-12), 83.9 (C-13), 80.5 (C-14), 143.0 (C-15), 113.5(C-16), 18.6 (C-17), 18.3 (C-18), 9.8 (C-19), 64.8 (C-20), 118.9 (C-1′),1′-Me: 21.5, 12-OBz: 165.4, 129.7, 129.4×2, 128.6×2, 133.3; HRESIMS m/z541.2070 [M+H]⁺ (calcd for C₂₉H₃₃O₁₀ ⁺, 541.2068).

Example 2

Compound YH-60 was obtained by isolating and purifying with thesemi-preparative HPLC. The structure and data of Compound YH-60 were asfollows:

YH-60: [α]²⁵ _(D)-14.2 (c 0.148, CH₂Cl₂); UV (MeOH) λ_(max) (log ε) 232(4.50) nm; IR (KBr) v_(max) 3418, 2925, 2855, 1704, 1631, 1452, 1379,1315, 1270, 1178, 1108, 1070, 1026, 1010, 940, 915 cm⁻¹; ¹H NMR (CDCl₃,400 MHz) δ_(H) 7.63 (1H, s, H-1), 4.13 (1H, s, H-5), 3.36 (1H, s, H-7),4.22 (1H, d, J=5.2 Hz, H-8), 3.51 (1H, s, H-10), 2.51 (1H, m, H-11),5.22 (1H, d, J=2.2 Hz, H-12), 6.19 (1H, d, J=5.2 Hz, H-14), 5.04 (1H, s,H-16a), 5.34 (1H, s, H-16b), 1.88 (3H, s, H₃-17), 1.40 (3H, d, J=7.3 Hz,H₃-18), 1.75 (3H, br s, H₃-19), 3.34 (1H, d, J=12.3 Hz, H-20a), 4.00(1H, d, J=12.3 Hz, H-20b), 12-OBz: 7.97 (2H, d, J=7.4 Hz), 7.49 (2H, m),7.60 (1H, m); 14-OBz: 8.07 (2H, d, J=7.0 Hz), 7.28 (2H, m), 7.60 (1H,m); ¹³C NMR (CDCl₃, 100 MHz) δ_(C) 159.6 (C-1), 137.0 (C-2), 208.6(C-3), 72.4 (C-4), 69.5 (C-5), 63.1 (C-6), 62.8 (C-7), 40.3 (C-8), 75.8(C-9), 51.0 (C-10), 42.9 (C-11), 79.9 (C-12), 75.2 (C-13), 73.7 (C-14),144.5 (C-15), 114.9 (C-16), 19.6 (C-17), 16.2 (C-18), 9.9 (C-19), 65.5(C-20), 12-OBz: 165.8, 129.4, 129.6×2, 128.6×2, 133.3, 14-OBz:166.6,129.8, 130.0×2, 128.4×2, 133.4; HRESIMS m/z 619.2168 [M−H]⁻ (calcd forC₃₄H₃₅O₁₁ ⁻, 619.2185).

Preparation of Derivatives

The raw material compounds used in the following examples were YH-6,YH-11, YH-16 and YH-22. The structures of these compounds were asfollows:

Example 3: Preparation of YH-7 and YH-8

Compound YH-6 (30 mg) was taken and dissolved in 2 mL of pyridine, andthen stirred under the protection of N₂. One-hundred (100) μL of aceticanhydride was pumped into with an injector and heated for reaction at50° C. Two products were found in thin layer detection. When the rawmaterial was basically reacted, 3 mL of water was added to stop thereaction, and then EtOAc (5 mL) was used to extract three times. Afterthe reaction product was purified with a preparative thin layer(CH₂Cl₂/MeOH, 50:1), YH-7 (10 mg) and YH-8 (15 mg) were obtained. Thestructure and data were as follows:

YH-7: UV (MeOH) λ_(max) (log) 240 (3.70) nm; ¹HNMR (400 MHz, CDCl₃)δ_(H) 7.72 (2H, m), 7.49 (1H, br s), 7.39 (3H, m), 5.57 (1H, s), 5.05(2H, s, overlap), 5.02 (1H, s), 4.91 (1H, d, J=2.5 Hz), 4.80 (1H, d,J=12.0 Hz), 4.06 (1H, m), 3.67 (1H, d, J=2.5 Hz), 3.63 (1H, d, J=12.0Hz), 3.53 (1H, s), 3.07 (1H, s), 2.37 (1H, q, J=7.3 Hz), 2.20 (3H, s),2.03 (6H, s), 1.88 (3H, s), 1.75 (1H, br s), 1.31 (3H, d, J=7.3 Hz); ¹³CNMR (100 MHz, CDCl₃) δ_(C) 10.0, 18.1, 18.8, 20.6, 20.7, 21.1, 35.4,43.9, 47.9, 59.5, 64.2, 66.3, 68.4, 71.8, 78.1, 78.3, 80.5, 84.1, 113.5,117.9, 126.0, 128.0, 129.7, 135.1, 137.1, 143.0, 158.2, 168.8, 169.6,170.6, 205.6; MS m/z 625.2 [M+H]⁺ 659.2 [M+Cl]⁻.

YH-8: [α]_(D) ²⁵+12.0 (c 0.47, MeOH), UV (MeOH) λ_(max) (log) 241 (4.16)nm; IR (KBr) v_(max) 3469, 2929, 1738, 1698, 1234, 1082, and 1024 cm⁻¹;¹H NMR (CDCl₃, 400 MHz) δ_(H) 7.57 (1H, br s, H-1), 4.30 (1H, s, H-5),3.51 (1H, s, H-7), 3.61 (1H, d, J=2.6 Hz, H-8), 3.96 (1H, m, H-10), 2.46(1H, q, J=7.3 Hz, H-11), 5.07 (1H, s, H-12), 4.88 (1H, d, J=2.6 Hz,H-14), 5.04 (1H, s, H-16a), 5.02 (1H, s, H-16b), 1.88 (3H, s, H₃-17),1.35 (3H, d, J=7.3 Hz, H₃-18), 1.78 (3H, brs, H₃-19), 4.83 (1H, d,J=12.0 Hz, H-20a), 3.92 (1H, d, J=12.0 Hz, H-20b). 1′-Ph: 7.71 (2H, m),7.38 (2H, m), 7.38 (1H, m), 12-OAc: 2.02 (3H. s). 20-OAc: 2.09 (3H, s);¹³C NMR (CDCl₃, 100 MHz) δ_(C) 160.1 (C-1), 136.9 (C-2), 209.2 (C-3),72.2 (C-4), 69.8 (C-5), 59.4 (C-6), 64.0 (C-7), 35.3 (C-8), 78.6 (C-9),47.3 (C-10), 44.0 (C-11), 78.2 (C-12), 84.0 (C-13), 80.7 (C-14), 143.0(C-15), 113.5 (C-16), 18.8 (C-17), 18.3 (C-18), 9.9 (C-19), 65.7 (C-20),117.9 (C-1′). 1′-Ph: 135.2, 126.0×2, 128.0×2, 129.6. 12-OAc: 169.6,21.1. 20-OAc: 170.6, 20.8; HRESIMS m/z 583.2179 [M+H]⁺ (calcd forC₃₁H₃₅O₁₁ ⁺, 583.2174).

Example 4: Preparation of Compound YH-9

Compound YH-6 (20 mg) was taken and dissolved in 2 mL ofdichloromethane. One-hundred (100) L of triethylamine (Et₃N) was addedunder stirring, and 100 L of benzoyl chloride was added for reaction for30 min. When the raw material was reacted, 5 mL of H₂O was added to stopthe reaction, and then dichloromethane (3×5 mL) was used to extract. Theorganic phase was concentrated, then purified with a gel (SephadexLH-20, MeOH) and a preparative thin layer (CH₂Cl₂/MeOH, 50:1), andfinally Compound YH-9 (11 mg) was obtained. The structure and data ofthe compound were as follows:

YH-9: [α]_(D) ²⁵+24 (c 0.43, MeOH); UV (MeOH) λ_(max) (log) 231 (3.50)nm; IR (KBr) v_(max) 3465, 2925, 1721, 1274, 1240, 1082, and 1023 cm⁻¹;¹H NMR (CDCl₃, 400 MHz) δ_(H) 7.58 (1H, br s, H-1), 4.39 (1H, s, H-5),3.59 (1H, s, H-7), 3.66 (1H, d, J=2.3 Hz, H-8), 4.01 (1H, br s, H-10),2.49 (1H, q, J=7.3 Hz, H-11), 5.08 (1H, s, H-12), 4.90 (1H, d, J=2.3 Hz,H-14), 5.03 (1H, s, H-16a), 5.05 (1H, s, H-16b), 1.88 (3H, s, H₃-17),1.37 (3H, d, J=7.3 Hz, H₃-18), 1.79 (3H, br s, H₃-19), 5.13 (1H, d,J=11.9 Hz, H-20a), 4.10 (1H, d, J=11.9 Hz, H-20b). 1′-Ph: 7.71 (2H, m),7.38 (2H, m), 7.38 (1H, m). 12-OAc: 2.02 (3H. s). 20-OBz: 8.05 (2H, d,J=7.7 Hz), 7.43 (2H, t, J=7.7 Hz), 7.56 (1H, m); ¹³C NMR (CDCl₃, 100MHz) δ_(C) 160.1 (C-1), 136.9 (C-2), 209.2 (C-3), 72.3 (C-4), 69.8(C-5), 59.7 (C-6), 64.2 (C-7), 35.3 (C-8), 78.6 (C-9), 47.3 (C-10), 44.0(C-11), 78.3 (C-12), 84.1 (C-13), 80.7 (C-14), 143.0 (C-15), 113.5(C-16), 18.8 (C-17), 18.3 (C-18), 9.9 (C-19), 66.6 (C-20), 117.9 (C-1′).1′-Ph: 135.2, 126.0×2, 128.0×2, 129.6. 12-OAc: 169.6, 21.1. 20-OBz:166.2, 129.8, 129.7×2, 128.4×2, 133.2; ESIMS m/z 645.2 [M+H]⁺, 679.2[M+Cl]⁻; HRESIMS m/z 643.2184 [M−H]⁻ (calcd for C₃₆H₃₅O₁₁ ⁻, 643.2185).

Example 5: Preparation of Compound YH-10

Compound YH-6 (20 mg) was taken and dissolved in anhydrous pyridine.Nitrogen was then added for protection after vacuuming, and 100 μL of2-thiophenecarbonyl chloride was pumped into with an injector and heatedfor reaction at 50° C. for 2 h. After the completion of the reaction, 5mL of water was added to quench, and then EtOAc (3×5 mL) was used toextract. The organic phase was concentrated, then purified with apreparative thin layer (CH₂Cl₂/MeOH, 50:1) and a gel (Sephadex LH-20,MeOH), and finally Compound YH-10 (12 mg) was obtained. The structureand data of the compound were as follows:

YH-10: Colorless oil: [α]_(D) ²⁵+17.1 (c 0.23, MeOH); UV (MeOH) λ_(max)(log) 245 (3.94) nm; IR (KBr) v_(max) 3476, 2924, 1706, 1257, 1230,1081, and 750 cm⁻¹; ¹H NMR (CDCl₃, 400 MHz) δ_(H) 7.57 (1H, br s, H-1),4.36 (1H, s, H-5), 3.57 (1H, s, H-7), 3.64 (1H, d, J=2.5 Hz, H-8), 3.98(1H, m, H-10), 2.48 (1H, q, J=7.2 Hz, H-11), 5.07 (1H, s, H-12), 4.90(1H, d, J=2.5 Hz, H-14), 5.05 (1H, s, H-16a), 5.02 (1H, s, H-16b), 1.88(3H, s, H₃-17), 1.36 (3H, d, J=7.2 Hz, H₃-18), 1.79 (3H, br s, H₃-19),5.12 (1H, d, J=11.9 Hz, H-20a), 4.05 (1H, d, J=11.9 Hz, H-20b). 1′-Ph:7.71 (2H, m), 7.38 (2H, m), 7.38 (1H, m). 12-OAc: 2.02 (3H. s).2-thenoyl: 7.82 (1H, dd, J=3.7, 1.1 Hz), 7.09 (1H, dd, J=4.8, 3.9 Hz),7.55 (1H, dd, J=4.8, 1.1 Hz); ¹³C NMR (CDCl₃, 100 MHz) δ_(C) 160.2(C-1), 137.0 (C-2), 209.2 (C-3), 72.3 (C-4), 69.7 (C-5), 59.7 (C-6),64.0 (C-7), 35.3 (C-8), 78.6 (C-9), 47.3 (C-10), 44.0 (C-11), 78.2(C-12), 84.1 (C-13), 80.7 (C-14), 143.0 (C-15), 113.6 (C-16), 18.9(C-17), 18.3 (C-18), 10.0 (C-19), 66.6 (C-20), 117.9 (C-1′). 1′-Ph:135.2, 126.0×2, 128.1×2, 129.7. 12-OAc: 169.7, 21.2. 2-thenoyl: 161.8,133.1, 133.9, 127.9, 132.8; ESIMS m/z 651.2 [M+H]⁺ [M+Cl]⁻; HRESIMS m/z651.1858 (calcd for C₃₄H₃₅O₁₁S⁺, 651.1855).

Example 6: Preparation of Compounds YH-12 and YH-13

Compound YH-11 (30 mg) was taken and dissolved in 2 mL of pyridine.Followed by stirring under the protection of N₂, 100 μL of aceticanhydride was pumped into with an injector and heated for reaction at50° C. Two products were found in thin layer detection. When the rawmaterial was basically reacted, 3 mL of water was added to stop thereaction, and EtOAc (5 mL) was used to extract three times. After thereaction products were purified with a preparative thin layer(CH₂Cl₂/MeOH, 50:1), YH-12 (13 mg) and YH-13 (10 mg) were obtained. Thestructure and data of the compounds were as follows:

YH-12: [α]²⁵+63.3 (c 0.33, MeOH); UV (MeOH) λ_(max) (log) 231 (3.99) nm;IR (KBr) v_(max) 3445, 2925, 1710, 1268, 1080, and 713 cm⁻¹; ¹H NMR(CDCl₃, 400 MHz) δ_(H) 7.50 (1H, br s, H-1), 5.55 (1H, s, H-5), 3.59(1H, s, H-7), 3.78 (1H, d, J=2.5 Hz, H-8), 4.08 (1H, m, H-10), 2.55 (1H,q, J=7.2 Hz, H-11), 5.28 (1H, s, H-12), 5.05 (1H, d, J=2.5 Hz, H-14),5.09 (1H, s, H-16a), 5.04 (1H, s, H-16b), 1.92 (3H, s, H₃-17), 1.40 (3H,d, J=7.2 Hz, H₃-18), 1.74 (3H, br s, H₃-19), 4.79 (1H, d, J=12.0 Hz,H-20a), 3.64 (1H, d, J=12.0 Hz, H-20b). 1′-Ph: 7.75 (2H, m), 7.40 (2H,m), 7.40 (1H, m). 12-OBz: 7.94 (2H, m), 7.48 (2H, m), 7.61 (1H, m).5-OAc: 2.14 (3H, s). 20-OAc: 2.03 (3H, s); ¹³C NMR (CDCl₃, 100 MHz)δ_(C) 158.0 (C-1), 137.2 (C-2), 205.4 (C-3), 71.7 (C-4), 68.4 (C-5),59.6 (C-6), 64.2 (C-7), 35.9 (C-8), 78.4 (C-9), 47.9 (C-10), 44.0(C-11), 78.9 (C-12), 84.3 (C-13), 80.6 (C-14), 142.9 (C-15), 113.8(C-16), 18.9 (C-17), 18.2 (C-18), 10.0 (C-19), 66.4 (C-20), 118.0(C-1′). 1′-Ph: 135.1, 126.0×2, 128.1×2, 129.7. 12-OBz: 165.4, 129.7,129.5×2, 128.6×2, 133.4. 5-OAc: 168.8, 20.7. 20-OAc: 170.6, 20.3; ESIMSm/z 687.3 [M+H]⁺; HRESIMS m/z 687.2420 [M+H]⁺ (calcd for C₃₈H₃₉O₁₂ ⁺,687.2436).

YH-13: white powder; [α]²⁵+50.2 (c 0.43, MeOH); UV (MeOH) λ_(max) (log)232 (4.02) nm; IR (KBr) v_(max) 3446, 2925, 1908, 1451, 1365, 1267,1241, 978, 713 cm⁻¹; ¹HNMR (CDCl₃, 400 MHz) δ_(H) 7.59 (1H, br s, H-1),4.28 (1H, d, J=2.4 Hz, H-5), 3.59 (1H, s, H-7), 3.73 (1H, d, J=2.5 Hz,H-8), 3.98 (1H, m, H-10), 2.64 (1H, q, J=7.3 Hz, H-11), 5.32 (1H, s,H-12), 5.01 (1H, d, J=2.5 Hz, H-14), 5.08 (1H, s, H-16a), 5.03 (1H, s,H-16b), 1.92 (3H, s, H₃-17), 1.45 (3H, d, J=7.3 Hz, H₃-18), 1.77 (3H, brs, H₃-19), 4.84 (1H, d, J=12.0 Hz, H-20a), 3.92 (1H, d, J=12.0 Hz,H-20b). 1′-Ph: 7.75 (2H, m), 7.40 (2H, m), 7.40 (1H, m). 12-OBz: 7.92(2H, m), 7.47 (2H, m), 7.59 (1H, m). 20-OAc: 2.09 (3H, s); ¹³C NMR(CDCl₃, 100 MHz) δ_(C) 160.1 (C-1), 136.9 (C-2), 209.2 (C-3), 72.1(C-4), 69.8 (C-5), 59.4 (C-6), 64.1 (C-7), 35.7 (C-8), 78.6 (C-9), 47.3(C-10), 44.1 (C-11), 78.9 (C-12), 84.2 (C-13), 80.7 (C-14), 142.9(C-15), 113.8 (C-16), 18.9 (C-17), 18.4 (C-18), 9.9 (C-19), 65.8 (C-20),118.0 (C-1′). 1′-Ph: 135.1, 126.0×2, 128.1×2, 129.7. 12-OBz: 165.4,129.7, 129.5×2, 128.6×2, 133.4. 20-OAc: 170.6, 20.9; ESIMS m/z 645.3[M+H]⁺; HRESIMS m/z 645.2339 [M+H]⁺ (calcd for C₃₆H₃₇O₁₁ ⁺, 645.2330).

Example 7: Preparation of Compound YH-25

Compound YH-22 (23 mg) was taken, and the product was prepared withreference to the method in Example 4. After the obtained product waspurified with a gel (Sephadex LH-20, MeOH) and a preparative thin layer(CH₂Cl₂/MeOH, 100:1), Compound YH-25 (15 mg) was obtained. The structureand data of the compound were as follows:

YH-25: Colorless oil; [α]_(D) ²⁵+28.5 (c 0.067, MeOH); UV (MeOH) λ_(max)(log) 230 (4.50) nm; IR (KBr) v_(max) 3459, 2926, 1719, 1273, 1230,1026, and 711 cm⁻¹; ¹HNMR (CDCl₃, 400 MHz) δ_(H) 7.57 (1H, m, H-1), 4.37(1H, s, H-5), 3.52 (1H, s, H-7), 3.57 (1H, d, J=2.5 Hz, H-8), 3.91 (1H,m, H-10), 2.40 (1H, q, J=7.2 Hz, H-11), 4.99 (1H, s, H-12), 4.76 (1H, d,J=2.5 Hz, H-14), 5.01 (1H, s, H-16a), 4.97 (1H, s, H-16b), 1.84 (3H, s,H₃-17), 1.30 (3H, d, J=7.2 Hz, H-18), 1.78 (3H, br s, H₃-19), 5.12 (1H,d, J=11.9 Hz, H-20a), 4.07 (1H, d, J=11.9 Hz, H-20b), 5.64 (1H, d,J=15.5 Hz, H-2′), 6.66 (1H, dd, J=15.5, 10.6 Hz, H-3′), 6.03 (1H, dd,J=15.0, 10.6 Hz, H-4′), 5.85 (1H, dd, J=15.0, 7.4 Hz, H-5′), 2.08 (2H,m, H₂-6′), 1.37 (2H, m, H₂-7′), 1.26 (2H, m, H₂-8′), 1.26 (2H, m,H₂-9′), 0.87 (3H, t, J=6.9 Hz, H-10′). 12-OAc: 1.99 (3H. s). 20-OBz:8.04 (2H, d, J=7.3 Hz), 7.44 (2H, m), 7.56 (1H, m); ¹³C NMR (CDCl₃, 100MHz) δ_(C) 160.2 (C-1), 136.9 (C-2), 209.2 (C-3), 72.3 (C-4), 69.7(C-5), 59.7 (C-6), 64.1 (C-7), 35.2 (C-8), 78.1 (C-9), 47.3 (C-10), 43.9(C-11), 78.2 (C-12), 83.6 (C-13), 80.3 (C-14), 143.1 (C-15), 113.3(C-16), 18.7 (C-17), 18.2 (C-18), 9.9 (C-19), 66.6 (C-20), 117.1 (C-1′),122.2 (C-2′), 135.0 (C-3′), 128.5 (C-4′), 139.3 (C-5′), 32.6 (C-6′),28.7 (C-7′), 31.3 (C-8′), 22.5 (C-9′), 14.0 (C-10′). 12-OAc: 169.7,21.1. 20-OBz: 166.2, 129.7, 129.7×2, 128.4×2, 133.1; ESIMS m/z 691.3[M+H]⁺, 725.2 [M+Cl]⁻; HRESIMS m/z 689.2962 [M−H]⁻ (calcd for C₃₉H₄₅O₁₁,689.2967).

Example 8: Preparation of Compound YH-33

Compound YH-22 (30 mg) was taken and dissolved in dichloromethane. Ten(10) mg of Grubbs second-generation catalyst was then added, andnitrogen was added for protection after vacuuming. Two hundred (200) μLof styrene was pumped into with an injector, and then heated forreaction under stirring at 40° C. for 1 h. The catalyst was removed byfiltration, the filtrate was concentrated and then purified with an HPLC(MeCN/H₂O, 75%, 3 mL/min), and finally Compound YH-33 (15 mg, t_(R)=10min) was obtained. The structure and data of the compound were asfollows:

YH-33: [α]_(D) ²⁵+4.2 (c 0.33, MeOH); UV (MeOH) λ_(max) (log) 286 (4.26)nm; ¹H NMR (CDCl₃, 400 MHz) δ_(H) 7.59 (1H, br s, H-1), 4.27 (1H, s,H-5), 3.57 (1H, s, H-7), 3.53 (1H, d, J=2.5 Hz, H-8), 3.85 (1H, m,H-10), 2.39 (1H, q, J=7.3 Hz, H-11), 5.00 (1H, s, H-12), 4.79 (1H, d,J=2.5 Hz, H-14), 5.03 (1H, br s, H-16a), 4.97 (1H, br s, H-16b), 1.85(3H, s, H₃-17), 1.31 (3H, d, J=7.3 Hz, H₃-18), 1.80 (3H, br s, H₃-19),3.94 (1H, d, J=12.4 Hz, H-20a), 3.81 (1H, d, J=12.4 Hz, H-20b), 5.88(1H, d, J=14.9 Hz, H-2′), 6.86 (1H, dd, J=14.9, 10.0 Hz, H-3′), 6.77(1H, dd, J=15.1, 10.0 Hz, H-4′), 6.69 (1H, d, J=15.1 Hz, H-5′), 7.40(2H, m, H-7′/H-11′), 7.32 (1H, dd, J=7.5, 7.5 Hz, H-8′/H-10′), 7.26 (1H,m, H-9′); 12-OAc: 2.00 (3H, s); ¹³C NMR (CDCl₃, 100 MHz) δ_(C) 160.3(C-1), 136.9 (C-2), 209.4 (C-3), 72.2 (C-4), 71.9 (C-5), 60.5 (C-6),64.2 (C-7), 35.4 (C-8), 78.3 (C-9), 47.4 (C-10), 44.0 (C-11), 78.2(C-12), 83.8 (C-13), 80.5 (C-14), 143.0 (C-15), 113.4 (C-16), 18.7(C-17), 18.3 (C-18), 9.9 (C-19), 65.1 (C-20), 116.8 (C-1′), 124.9(C-2′), 134.8 (C-3′), 127.0 (C-4′), 136.0 (C-5′), 136.7 (C-6′), 126.7(C-7′/C-11′), 128.6 (C-8′/10′), 128.1 (C-9′), 12-OAc: 169.7, 21.2;HRESIMS m/z 615.2206 [M+Na]⁺ (calcd for, C₃₃H₃₆O₁₀Na+, 615.2201) and627.1993 [M+Cl]⁻ (calcd for C₃₃H₃₆O₁₀Cl⁻, 627.2002).

Example 9: Preparation of Compound YH-34

Compound YH-16 (30 mg) was taken, and the product was prepared withreference to the method in Example 8. After the obtained product waspurified with an HPLC (MeCN/H₂O, 80%, 3 mL/min), Compound YH-34 (16 mg,t_(R)=14 min) was obtained. The structure and data of the compound wereas follows:

YH-34: [α]²⁵+27.8 (c 0.13, MeOH); UV (MeOH) λ_(max) (log) 285 (4.24) nm;¹H NMR (CDCl₃, 500 MHz) δ_(H) 7.61 (1H, br s, H-1), 4.23 (1H, s, H-5),3.66 (1H, s, H-7), 3.65 (1H, d, J=2.7 Hz, H-8), 3.87 (1H, m, H-10), 2.58(1H, q, J=7.2 Hz, H-11), 5.24 (1H, s, H-12), 4.93 (1H, d, J=2.7 Hz,H-14), 5.04 (1H, br s, H-16a), 5.02 (1H, br s, H-16b), 1.89 (3H, s,H₃-17), 1.41 (3H, d, J=7.2 Hz, H₃-18), 1.79 (3H, br s, H₃-19), 3.94 (1H,d, J=12.2 Hz, H-20a), 3.83 (1H, d, J=12.2 Hz, H-20b), 5.91 (1H, d,J=15.2 Hz, H-2′), 6.90 (1H, dd, J=15.2, 10.5 Hz, H-3′), 6.78 (1H, dd,J=15.4, 10.5 Hz, H-4′), 6.73 (1H, d, J=15.4 Hz, H-5′), 7.41 (2H, d,J=7.6 Hz, H-7′/H-11′), 7.33 (1H, dd, J=7.6, 7.6 Hz, H-8′/H-10′), 7.26(1H, m, H-9′); 12-OBz: 7.90 (2H, d, J=7.6 Hz), 7.46 (2H, dd, J=7.6, 7.6Hz), 7.58 (1H, dd, J=7.6, 7.6 Hz); ¹³C NMR (CDCl₃, 125 MHz) δ_(C) 160.2(C-1), 137.0 (C-2), 209.3 (C-3), 72.2 (C-4), 71.8 (C-5), 60.6 (C-6),64.0 (C-7), 35.8 (C-8), 78.4 (C-9), 47.4 (C-10), 44.1 (C-11), 78.9(C-12), 84.0 (C-13), 80.5 (C-14), 142.9 (C-15), 113.7 (C-16), 18.8(C-17), 18.3 (C-18), 9.9 (C-19), 64.7 (C-20), 116.9 (C-1′), 124.8(C-2′), 134.8 (C-3′), 127.0 (C-4′), 136.1 (C-5′), 136.7 (C-6′), 126.7(C-7′/C-11′), 128.6 (C-8′/10′), 128.1 (C-9′), 12-OBz: 129.6, 129.5×2,128.7×2, 133.3; HRESIMS m/z 689.2147 [M+Cl]⁻ (calcd for C₃₈H₃₈O₁₀Cl⁻,689.2159).

Example 10: Preparation of Compound YH-37

Compound YH-22 (20 mg) was taken and dissolved in 3 mL of methanol.Twenty (20) mg of palladium-carbon catalyst was then added, and hydrogenwas added for protection after vacuuming. Followed by stirring forreaction at a room temperature for 30 min, the palladium-carbon wasremoved by filtration. The filtrate was concentrated, then purifieddirectly with an HPLC (MeCN/H₂O, 95%, 3 mL/min), and finally CompoundYH-37 (12 mg, t_(R)=14 min) was obtained. The structure and data of thecompound were as follows:

YH-37: [α]²⁵+87.0 (c 0.033, MeOH); UV (MeOH) λ_(max) (log) 243 (4.03)nm; IR (KBr) v_(max) 3465, 2925, 1744, 1698, 1230, 1027, and 936 cm⁻¹;¹H NMR (CDCl₃, 400 MHz) δ_(H) 7.53 (1H, br s, H-1), 4.23 (1H, s, H-5),3.49 (1H, s, H-7), 3.34 (1H, d, J=2.4 Hz, H-8), 3.73 (1H, m, H-10), 2.30(1H, q, J=7.3 Hz, H-11), 4.93 (1H, s, H-12), 4.44 (1H, d, J=2.4 Hz,H-14), 1.85 (1H, m, H-15), 0.91 (3H, d, J=6.9 Hz, H₃-16), 0.93 (3H, d,J=6.9 Hz, H₃-17), 1.22 (3H, d, J=7.3 Hz, H₃-18), 1.78 (3H, br s, H₃-19),3.89 (1H, d, J=12.4 Hz, H-20a), 3.79 (1H, d, J=12.4 Hz, H-20b), 1.87(2H, m, H₂-2′), 1.54 (2H, m, H₂-3′), 1.27-1.30 (8H, m, H-4′-H-7′), 1.30(2H, m, H₂-8′), 1.26 (2H, m, H₂-9′), 0.87 (3H, t, J=6.8 Hz, H-10′);12-OAc: 2.00 (3H, s); ¹³C NMR (CDCl₃, 100 MHz) δ_(C) 160.9 (C-1), 136.6(C-2), 209.7 (C-3), 72.2 (C-4), 72.0 (C-5), 60.4 (C-6), 64.5 (C-7), 35.5(C-8), 77.2 (C-9), 47.5 (C-10), 44.0 (C-11), 77.5 (C-12), 83.6 (C-13),80.0 (C-14), 31.3 (C-15), 16.6 (C-16), 15.9 (C-17), 18.1 (C-18), 9.8(C-19), 65.2 (C-20), 119.9 (C-1′), 34.8 (C-2′), 23.4 (C-3′), 29.6(C-4′), 29.5 (C-5′), 29.5 (C-6′), 29.3 (C-7′), 22.7 (C-8′), 31.8 (C-9′),14.1 (C-10′); 12-OAc: 169.9, 21.2; HRESIMS m/z 593.3315 [M+H]⁺ (calcdfor C₃₂H₄₉O₁₀ ⁺, 593.3320).

Example 11: Preparation of Compound YH-38

Compound YH-22 (20 mg) was taken and dissolved in 3 mL of methanol.Thirty (30) mg of palladium-carbon catalyst was then added, and hydrogenwas added for protection after vacuuming. Followed by stirring forreaction at 50° C. for 30 min, the palladium-carbon was removed byfiltration. The filtrate was concentrated, and then purified with anHPLC (MeCN/H₂O, 95%, 3 mL/min), and finally Compound YH-38 (9 mg,t_(R)=15 min) was obtained. The structure and data of the compound wereas follows:

YH-38: [α]²⁵+76.8 (c 0.67, MeOH); UV (MeOH) λ_(max) (log) 202 (3.50) nm;IR (KBr) v_(max) 3463, 2927, 1742, 1378, 1229, 1028, and 940 cm⁻¹; ¹HNMR (CDCl₃, 500 MHz) δ111.53 (1H, m, H-la), 2.28 (1H, m, H-1b) 2.28 (1H,m, H-2), 4.07 (1H, s, H-5), 3.44 (1H, s, H-7), 3.32 (1H, s, H-8), 2.84(1H, m, H-10), 2.25 (1H, m, H-11), 5.03 (1H, s, H-12), 4.40 (1H, s,H-14), 1.83 (1H, m, H-15), 0.93 (3H, d, J=6.7 Hz, H₃-16), 0.93 (3H, d,J=6.7 Hz, H₃-17), 1.25 (3H, d, J=7.3 Hz, H₃-18), 1.10 (3H, d, J=5.5 Hz,H₃-19), 3.87 (1H, d, J=12.2 Hz, H-20a), 3.75 (1H, d, J=12.2 Hz, H-20b),1.84 (2H, m, H₂-2′), 1.52 (2H, m, H₂-3′), 1.25-1.31 (8H, m, H-4′-H-7′),1.28 (2H, m, H₂-8′), 1.26 (2H, m, H₂-9′), 0.87 (3H, t, J=6.7 Hz, H-10′);12-OAc: 2.00 (3H, s); ¹³C NMR (CDCl₃, 125 MHz) δ_(C) 33.3 (C-1), 42.8(C-2), 220.3 (C-3), 75.0 (C-4), 71.1 (C-5), 60.7 (C-6), 64.5 (C-7), 35.5(C-8), 77.6 (C-9), 44.0 (C-10), 43.5 (C-11), 76.8 (C-12), 83.1 (C-13),80.3 (C-14), 31.2 (C-15), 16.5 (C-16), 15.9 (C-17), 18.3 (C-18), 12.4(C-19), 65.4 (C-20), 120.0 (C-1′), 34.9 (C-2′), 23.3 (C-3′), 29.6(C-4′), 29.5 (C-5′), 29.5 (C-6′), 29.3 (C-7′), 22.6 (C-8′), 31.8 (C-9′),14.1 (C-10′); 12-OAc: 170.0, 21.2; HRESIMS m/z 617.3316 [M+Na]⁺ (calcdfor C₃₂H₅₀O₁₀ ⁺, 617.3296).

Example 12: Preparation of Compound YH-39

Compound YH-11 was taken, and the product was prepared with reference tothe method in Example 11. After the obtained product was purified withan HPLC (MeCN/H₂O, 70%, 3 mL/min), Compound YH-39 (10 mg, t_(R)=17 min)was obtained. The structure and data of the compound were as follows:

YH-39: [α]²⁵+64.5 (c 0.067, MeOH); UV (MeOH) λ_(max) (log) 230 (4.08)nm; IR (KBr) v_(max) 3483, 2968, 1720, 1272, 1079, and 1025 cm⁻¹; ¹H NMR(CDCl₃, 400 MHz) δ112.36 (1H, m, H-1α), 1.61 (1H, m, H-1β), 2.25 (1H, m,H-2), 4.05 (1H, s, H-5), 3.62 (1H, s, H-7), 3.59 (1H, d, J=2.4 Hz, H-8),3.01 (1H, dd, J=13.2, 5.8 Hz, H-10), 2.54 (1H, q, J=6.9 Hz, H-11), 5.42(1H, s, H-12), 4.74 (1H, d, J=2.4 Hz, H-14), 2.03 (1H, m, H-15), 1.04(3H, d, J=6.7 Hz, H₃-16), 1.04 (3H, d, J=6.7 Hz, H₃-17), 1.46 (3H, d,J=6.9 Hz, H₃-18), 1.09 (3H, d, J=6.6 Hz, H₃-19), 3.86 (1H, d, J=12.3 Hz,H-20a), 3.79 (1H, d, J=12.3 Hz, H-20b); 1′-Ph: 7.75 (2H, m), 7.39 (2H,m), 7.39 (1H, m). 12-OBz: 7.94 (2H, d, J=7.4 Hz), 7.47 (2H, dd, J=7.4,7.4 Hz), 7.59 (1H, dd, J=7.4, 7.4 Hz); ¹³C NMR (CDCl₃, 100 MHz) δ_(C)33.4 (C-1), 42.8 (C-2), 220.1 (C-3), 74.9 (C-4), 71.2 (C-5), 60.7 (C-6),64.4 (C-7), 36.1 (C-8), 78.9 (C-9), 44.1 (C-10), 43.8 (C-11), 77.3(C-12), 84.1 (C-13), 81.0 (C-14), 31.6 (C-15), 16.7 (C-16), 16.2 (C-17),18.6 (C-18), 12.4 (C-19), 65.1 (C-20), 118.1 (C-1′). 1′-Ph: 135.7,125.9×2, 128.0×2, 129.5. 12-OBz: 165.6, 129.8, 129.5×2, 128.6×2, 133.3;HRESIMS m/z 605.2407 [M−H](calcd for C₃₄H₃₇O₁₀ ⁻, 605.2392).

Example 13: Preparation of Compound YH-47 and Compound YH-48

Compound YH-6 (30 mg) was taken and dissolved in 2 mL of tetrahydrofuran(THF). Two-hundred (200) μL of concentrated hydrochloric acid was addedfor reaction for about 20 min under stirring, 10 mL of water was addedto stop the reaction, and then EtOAc (3×10 mL) was added for extraction.After the reaction products passed through a gel (MeOH) and apreparative thin layer (CH₂Cl₂/MeOH, 40:1), Compound YH-47 (13 mg) andCompound YH-48 (10 mg) were obtained. The structure and data of thecompounds were as follows:

YH-47:[α]_(D) ²⁵+10.5 (c 0.13,MeOH); UV (MeOH) λ_(max) (log) 241 (3.70)nm; IR (KBr) v_(max) 3451, 2925, 1740, 1691, 1230, 1078, and 697 cm⁻¹;¹H NMR (CDCl₃, 400 MHz) δ_(H) 7.61 (1H, br s, H-1), 4.38 (1H, s, H-5),4.74 (1H, s, H-7), 3.60 (1H, d, J=2.4 Hz, H-8), 4.04 (1H, m, H-10), 2.85(1H, q, J=7.3 Hz, H-11), 5.03 (1H, s, H-12), 4.96 (1H, d, J=2.4 Hz,H-14), 5.03 (1H, s, H-16a), 5.06 (1H, s, H-16b), 1.87 (3H, s, H₃-17),1.35 (3H, d, J=7.3 Hz, H₃-18), 1.81 (3H, br s, H₃-19), 4.01 (1H, d,J=11.3 Hz, H-20a), 4.16 (1H, d, J=11.3 Hz, H-20b). 1′-Ph: 7.64 (2H, m),7.41 (2H, m), 7.40 (1H, m). 12-OAc: 1.98 (3H. s); ¹³C NMR (CDCl₃, 100MHz) δ_(C) 158.6 (C-1), 136.7 (C-2), 208.6 (C-3), 75.2 (C-4), 70.9(C-5), 76.9 (C-6), 81.4 (C-7), 36.2 (C-8), 78.9 (C-9), 49.9 (C-10), 43.5(C-11), 77.7 (C-12), 84.7 (C-13), 82.7 (C-14), 142.4 (C-15), 113.9(C-16), 18.7 (C-17), 17.8 (C-18), 9.9 (C-19), 68.8 (C-20), 117.7 (C-1′).1′-Ph: 134.6, 125.8×2, 128.2×2, 130.0. 12-OAc: 169.5, 20.9; HRESIMS m/z575.1685 [M−H](calcd. for C₂₉H₃₂O₁₀Cl⁻, 575.1689).

YH-48:[α]_(D) ²⁵+15 (c 0.1, MeOH); UV (MeOH) λ_(max) (log) 241 (3.70)nm; IR (KBr) v_(max) 3451, 2925, 1740, 1691, 1230, 1078, and 697 cm⁻¹;¹H NMR (CDCl₃, 400 MHz) δ_(H) 7.59 (1H, br s, H-1), 3.96 (1H, s, H-5),4.89 (1H, d, J=9.8 Hz, H-7), 3.06 (1H, m, H-8), 3.07 (1H, m, H-10), 2.85(1H, q, J=7.2 Hz, H-11), 5.02 (1H, s, H-12), 5.37 (1H, d, J=2.4 Hz,H-14), 5.01 (1H, s, H-16a), 5.03 (1H, s, H-16b), 1.85 (3H, s, H₃-17),1.33 (3H, d, J=7.2 Hz, H₃-18), 1.80 (3H, br s, H₃-19), 4.06 (1H, d,J=10.4 Hz, H-20a), 4.46 (1H, d, J=10.4 Hz, H-20b). 1′-Ph: 7.67 (2H, m),7.39 (2H, m), 7.39 (1H, m). 12-OAc: 2.03 (3H. s); ¹³C NMR (CDCl₃, 100MHz) δ_(C) 158.9 (C-1), 137.6 (C-2), 208.5 (C-3), 73.3 (C-4), 81.7(C-5), 79.0 (C-6), 66.9 (C-7), 36.8 (C-8), 78.4 (C-9), 50.6 (C-10), 42.8(C-11), 77.9 (C-12), 84.2 (C-13), 79.1 (C-14), 142.8 (C-15), 113.7(C-16), 18.8 (C-17), 18.1 (C-18), 10.0 (C-19), 61.8 (C-20), 117.4(C-1′). 1′-Ph: 134.8, 125.9×2, 128.1×2, 129.8. 12-OAc: 169.7, 21.0;HRESIMS m/z 575.1681 [M−H]⁻ (calcd. for C₂₉H₃₂O₁₀Cl⁻, 575.1689).

Example 14: Preparation of Compound YH-49 and Compound YH-50

Compound YH-11 (40 mg) was taken, and the product was prepared by usingthe same method in Example 13. After purifying the obtained product witha preparative thin layer (CH₂Cl₂/MeOH, 50:1), Compound YH-49 (20 mg) andCompound YH-50 (12 mg) were obtained. The structure and data of thecompounds were as follows:

YH-49:[α]_(D) ²⁵+23.6 (c 0.46,MeOH); UV (MeOH) λ_(max) (log) 231 (4.01)nm; IR (KBr) v_(max) 3450, 2924, 1721, 1268, 1079, and 711 cm⁻¹; ¹H NMR(CDCl₃, 400 MHz) δ_(H) 7.64 (1H, s, H-1), 4.40 (1H, s, H-5), 4.84 (1H,s, H-7), 3.79 (1H, d, J=2.4 Hz, H-8), 4.08 (1H, m, H-10), 2.99 (1H, q,J=7.3 Hz, H-11), 5.34 (1H, s, H-12), 5.10 (1H, d, J=2.4 Hz, H-14), 5.07(1H, s, H-16a), 5.03 (1H, s, H-16b), 1.91 (3H, s, H₃-17), 1.45 (3H, d,J=7.3 Hz, H₃-18), 1.81 (3H, br s, H₃-19), 4.16 (1H, d, J=11.5 Hz,H-20a), 4.02 (1H, d, J=11.5 Hz, H-20b). 1′-Ph: 7.68 (2H, m), 7.42 (2H,m), 7.44 (1H, m). 12-OBz: 7.97 (2H, m), 7.42 (2H, m), 7.55 (1H, t, J=7.4Hz); ¹³C NMR (CDCl₃, 100 MHz) δ_(C) 158.6 (C-1), 136.9 (C-2), 208.6(C-3), 75.1 (C-4), 70.9 (C-5), 77.0 (C-6), 81.6 (C-7), 36.6 (C-8), 79.1(C-9), 49.9 (C-10), 43.8 (C-11), 78.0 (C-12), 85.1 (C-13), 82.9 (C-14),142.4 (C-15), 114.2 (C-16), 18.8 (C-17), 18.0 (C-18), 9.9 (C-19), 68.9(C-20), 117.9 (C-1′). 1′-Ph: 134.6, 125.8×2, 128.3×2, 130.1. 12-OBz:165.1, 129.4, 129.6×2, 128.4×2, 133.4; HRESIMS m/z 637.18513 [M−H]⁻(calcd for C₃₄H₃₄O₁₀Cl⁻, 637.18460).

YH-50: [α]_(D) ²⁵+16.7 (c 0.35, MeOH); UV (MeOH) λ_(max) (log) 230(4.10) nm; IR (KBr) v_(max) 3431, 2960, 1721, 1451, 1268, and 1075 cm⁻¹;¹H NMR (CDCl₃, 500 MHz) δ_(H) 7.62 (1H, br s, H-1), 3.90 (1H, s, H-5),4.90 (1H, d, J=9.9 Hz, H-7), 3.12 (1H, d, J=9.9 Hz, H-8), 3.06 (1H, brs, H-10), 2.99 (1H, q, J=7.2 Hz, H-11), 5.28 (1H, s, H-12), 5.51 (1H, s,H-14), 5.06 (1H, s, H-16a), 5.01 (1H, s, H-16b), 1.88 (3H, s, H₃-17),1.44 (3H, d, J=7.2 Hz, H-18), 1.78 (3H, br s, H-19), 4.33 (1H, d, J=11.1Hz, H-20a), 4.06 (1H, d, J=11.1 Hz, H-20b). 1′-Ph: 7.71 (2H, d, J=7.3Hz), 7.42 (2H, m), 7.43 (1H, m). 12-OBz: 7.94 (2H, d, 7.4), 7.47 (2H,m), 7.59 (1H, t, J=7.4 Hz); ¹³C NMR (CDCl₃, 125 MHz) δ_(C) 158.9 (C-1),137.6 (C-2), 208.4 (C-3), 73.1 (C-4), 81.8 (C-5), 78.5 (C-6), 66.5(C-7), 37.1 (C-8), 78.4 (C-9), 50.4 (C-10), 43.0 (C-11), 78.5 (C-12),84.4 (C-13), 79.2 (C-14), 142.8 (C-15), 114.0 (C-16), 18.9 (C-17), 18.3(C-18), 10.0 (C-19), 62.1 (C-20), 117.5 (C-1′). 1′-Ph: 134.8, 125.9×2,128.2×2, 130.1. 12-OBz: 165.0, 129.4, 129.4×2, 128.7×2, 133.6; HRESIMSm/z 637.18536 [M−H]⁻ (calcd for C₃₄H₃₄O₁₀Cl⁻, 637.18460).

Example 15: Preparation of Compound YH-52 and Compound YH-53

Compound YH-11 (100 mg) was taken and dissolved in 3 mL ofdichloromethane, then stirred at −60° C. for 5 minutes, and PBr₃ (50 μL)was added. After the thin layer detects the end of the reaction, thereaction solution was taken out and then quenched by the addition of 5mL of water. Followed by adding dichloromethane for extraction (3×5 mL),the organic layer was combined, concentrated under a reduced pressure,then purified with a semi-preparative high-performance liquid phase(MeCN/H₂O, 80: 20, 3 mL/min), and finally Compound YH-52 (23 mg,t_(R)=15 min) and Compound YH-53 (20 mg, t_(R)=12 min) were obtained.The structure and data of the compounds were as follows:

YH-52: [α]_(D) ²⁵+56.3 (c 0.35, MeOH); UV (MeOH) λ_(max) (log) 232(3.96) nm; IR (KBr) v_(max) 3445, 2923, 1720, 1692, 1268, 1078, 1008,and 710 cm⁻¹; ¹H NMR (CDCl₃, 400 MHz) δ_(H) 7.64 (1H, s, H-1), 4.27 (1H,s, H-5), 4.98 (1H, s, H-7), 3.88 (1H, d, J=2.4 Hz, H-8), 4.08 (1H, s,H-10), 3.00 (1H, q, J=7.3 Hz, H-11), 5.33 (1H, s, H-12), 5.11 (1H, d,J=2.4 Hz, H-14), 5.04 (1H, s, H-16a), 5.08 (1H, s, H-16b), 1.91 (3H, s,H₃-17), 1.45 (3H, d, J=7.3 Hz, H₃-18), 1.81 (3H, br s, H₃-19), 4.10 (1H,d, J=12.0 Hz, H-20a), 4.21 (1H, m, H-20b), 12-OBz: 8.01 (2H, m), 7.39(2H, m), 7.55 (1H, m); 1′-Ph: 7.68 (2H, m), 7.44 (3H, m); ¹³C NMR(CDCl₃, 100 MHz) δ_(C) 158.6 (C-1), 136.9 (C-2), 208.7 (C-3), 75.1(C-4), 70.5 (C-5), 75.8 (C-6), 82.5 (C-7), 37.8 (C-8), 79.2 (C-9), 50.0(C-10), 43.8 (C-11), 78.1 (C-12), 85.1 (C-13), 82.8 (C-14), 142.4(C-15), 114.2 (C-16), 18.8 (C-17), 18.0 (C-18), 9.9 (C-19), 69.9 (C-20),117.9 (C-1′), 1′-Ph: 134.5, 125.8×2, 128.4×2, 130.1, 12-OBz: 165.1,129.3, 129.8×2, 128.3×2, 133.4; HRESIMS m/z 705.1309 [M+Na]⁺ (calcd for,C₃₄H₃₅O₁₀BrNa⁺, 705.1306).

YH-53: [α]²⁵+50.0 (c 0.30, MeOH); UV (MeOH) λ_(max) (log) 232 (4.06) nm;IR (KBr) v_(max) 3428, 1701, 1452, 1268, 1079, 1026, and 712 cm⁻¹; ¹HNMR (CDCl₃, 500 MHz) δ_(H) 7.57 (1H, s, H-1), 3.85 (1H, s, H-5), 5.06(1H, d, J=9.9 Hz, H-7), 3.12 (1H, d, J=9.9 Hz, H-8), 3.07 (1H, br s,H-10), 3.00 (1H, q, J=7.2 Hz, H-11), 5.28 (1H, s, H-12), 5.57 (1H, d,J=1.3 Hz, H-14), 5.00 (1H, s, H-16a), 5.04 (1H, s, H-16b), 1.88 (3H, s,H₃-17), 1.42 (3H, d, J=7.2 Hz, H₃-18), 1.73 (3H, br s, H₃-19), 4.09 (1H,d, J=10.9 Hz, H-20a), 4.39 (1H, d, J=10.9 Hz, H-20b), 12-OBz: 7.93 (2H,d, J=7.5 Hz), 7.46 (2H, dd, J=7.5, 7.5 Hz), 7.56 (1H, m); 1′-Ph: 7.71(2H, m), 7.42 (2H, m), 7.41 (1H, m); ¹³C NMR (CDCl₃, 125 MHz) δ_(C)158.8 (C-1), 137.5 (C-2), 208.2 (C-3), 73.4 (C-4), 81.1 (C-5), 78.2(C-6), 63.7 (C-7), 36.9 (C-8), 78.5 (C-9), 50.7 (C-10), 43.0 (C-11),78.4 (C-12), 84.7 (C-13), 81.8 (C-14), 142.7 (C-15), 114.0 (C-16), 18.9(C-17), 18.2 (C-18), 9.9 (C-19), 63.6 (C-20), 117.3 (C-1′), 1′-Ph:134.9, 125.9×2, 128.1×2, 129.8, 12-OBz: 165.0, 129.4, 129.4×2, 128.7×2,133.6; HRESIMS m/z 705.1289 [M+Na]⁺ (calcd for, C₃₄H₃₅O₁₀BrNa⁺,705.1306).

Inhibitory Activity of Daphnane Diterpenoid Compound onCastration-Resistant Prostate Cancer Cells (1) Cells Culture

All prostate cancer cells were cultured with a RPMI-1640 mediumcontaining 10% calf serum, 100 units of penicillin per milliliter and100 g/mL of streptomycin in a constant temperature incubator containing5% carbon dioxide at a saturated humidity of 37° C.

(2) Cytotoxic Activity Test

Cells in the logarithmic phase of growth were taken and cultured in96-well plates for 24 hours (5×10³ cells/well), then treated withdifferent concentrations of the compounds to be tested, and incubated byMTT method for 4 hours. The supernatant was removed by centrifugation,the MTT crystals were dissolved by adding DMSO, and the absorbance valuewas measured by an enzyme-linked immunosorbent assay at a wavelength of570 nm. The cytotoxic activity of the compounds to be tested on cancercells was expressed as IC₅₀.

(3) Experimental Results

As shown in Table 1, most of the daphnane diterpenoids showed differentdegrees of activity in inhibiting a variety of prostate cancer cells.Some representative compounds had a significant inhibitory activity onprostate cancer cells, with an IC₅₀ at an nM level, which were strongerthan that of the positive drugs doxorubicin and enzalutamide (ENZ),among which the activity of the compounds YH-11, YH-22 and YH-52 werethe most prominent. A preliminary structure-activity relationshipanalysis showed that the inhibitory activity of the compounds thatcontain 9,13,14-orthoesters (YH-11 and YH-52, etc.) was significantlyhigher than that of compounds without orthoesters (YH-56, YH-60 andYH-61), which indicated that the orthoester was the key pharmacophore.Next, the substitution of 20-OH by an ester-philic groups may lead to asignificant decrease in activity, such as the activity of the compoundsYH-5, YH-9, YH-12, YH-19, YH-20, YH-25, etc. Additionally, the 6,7-epoxyhad a significantly reduced activity when being ring-opened to form adihydroxy product YH-51, and had an enhanced activity and asignificantly reduced toxicity for the normal human prostate cell RWPE-1than that of prototype compound YH-11 when being ring-opened to form abromination product YH-52. Such compounds having modifications to the1′-olefin side chain had unforeseen effects on activity, but overall,compounds with 1′-olefin side chains were more cytotoxic, for example,the compounds YH-16 and YH-22 were more cytotoxic than compounds YH-11and YH-6. Compounds YH-52 was selected as a candidate for in vivopharmacodynamic evaluation in animals.

TABLE 1 inhibitory activity of representative daphnane diterpenoidscompounds on several human prostate cancer cells and normal cellsProstate cancer cells IC₅₀ (μM) Normal cells No. C4-2B C4-2B/ENZR LNcapVcap-CRPC 22RV1 RWPE-1 YH-1 3.41 × e⁻⁶ 1.56 × e⁻⁴ 3.76 × e⁻⁵ 1.41 × e⁻⁵0.00574 43.5 YH-2 8.21 × e⁻⁵ 3.36 × e⁻⁵ 4.25 × e⁻⁵ 1.17 × e⁻⁵ 0.026540.3 YH-3 0.00895 4.03 × e⁻⁴ 0.00377 0.00174 0.434 57.2 YH-5 0.904 0.4340.0993 4.66 44.5 72.8 YH-6 0.000367 0.00234 0.00389 0.0212 0.0525 44.2YH-9 0.380 1.462 NA NA NA NA YH-11 8.14 × e⁻⁷ 2.46 × e⁻⁷ 3.31 × e⁻⁷ 2.93× e⁻⁷ 1.36 × e⁻⁴ 13.4 YH-12 0.557 1.90 NA NA NA NA YH-13 7.10 × e⁻³0.00679 0.00453 0.0672 0.0832 35.7 YH-16 3.42 × e⁻⁴ 2.32 × e⁻⁴ 0.005430.00570 0.0332  9.67 YH-19 0.0644 0.136 NA NA NA NA YH-20 1.578 5.76 NANA NA NA YH-22 1.62 × e⁻⁸ 1.66 × e⁻⁷ 2.31 × e⁻⁶ 5.63 × e⁻⁷ 3.46 × e⁻⁴ 5.89 YH-24 1.97 × e⁻³ 0.0673 0.0482 0.0762 0.0458 21.9 YH-25 0.593 2.30NA NA NA NA YH-30 0.0219 0.0368 0.00248 0.171 7.23 73.5 YH-35 0.0004260.00192 0.00482 0.0642 0.144 42.9 YH-36 3.15 × e⁻⁴ 8.46 × e⁻⁵ 3.96 × e⁻⁴1.53 × e⁻⁶ 0.0425 14.6 YH-37 9.98 × e⁻⁸ 1.45 × e⁻⁷  2.22 e⁻⁵ 1.26 × e⁻⁴5.08 × e⁻⁴ 10.3 YH-38 1.56 × e⁻⁶ 1.23 × e⁻⁵ 2.76 × e⁻⁴ 9.02 × e⁻⁴0.00542 20.6 YH-39 2.99 × e⁻⁵ 2.32 × e⁻⁵ 2.98 × e⁻⁴ 7.90 × e⁻⁴ 0.0067318.5 YH-40 3.59 × e⁻⁶ 9.34 × e⁻⁵ 3.02 × e⁻⁵ 2.20 × e⁻⁴ 1.20 × e⁻⁴ 20.3YH-44 0.193 0.641 0.0748 0.306 4.65 17.4 YH-51 0.00781 0.0774 0.002510.00202 1.77 12.3 YH-52 5.17 × e⁻⁷ 4.66 × e⁻⁸ 4.22 × e⁻⁶ 5.93 × e⁻⁸ 9.67× e⁻⁵ 53.6 YH-53 2.35 × e⁻⁶ 5.77 × e⁻⁵ 3.56 × e⁻⁵ 4.33 × e⁻⁴ 1.98 × e⁻⁴56.3 YH-56 4.47 2.69 13.1 15.4 47.1 35.1 YH-57 2.32 1.87 4.82 40.8 46.741.1 YH-58 33.6 40.2 NA NA NA NA YH-60 0.0494 0.0739 0.00285 0.911 3.89 4.81 YH-61 0.189 0.543 NA NA NA NA ENZ 24.8 62.3 20.7 57.3 39.4 NA DOX0.00104 0.00287 2.55 × e⁻⁵ 0.0124 0.00599   0.0333 ENZ: a positive drugenzalutamide; DOX: a positive drug doxorubicin; and NA means not testedfor a low activity.

In Vivo Pharmacodynamic Evaluation of Compound YH-52 in Animals (1)Construction of a Mice Model for 22RV1 Prostate Cancer Growth

First, a large number of 22RV1 cells were amplified. To ensure thesuccess rate of tumor-bearing, the cell state needs to be adjusted tothe optimal state. When the number of the cells was sufficient, thecells were digested and flushed twice with PBS buffer to remove fetalbovine serum (FBS) from the medium to reduce immune rejection. The cellswere adjusted to 3000*10⁴/mL after counting by a hemocytometer. Then22Rv1 human prostate cancer cells (1004) were subcutaneously injectedinto a 4-5 weeks old nod-scid male mice with severe immunodeficiency.

(2) Grouping and Administration Oral Administration Experiment:

When the tumor grew to a volume of about 100 mm³, the mice were randomlydivided into 3 groups to make sure that the average tumor volume was thesame. The mice in the dosing group were orally gavaged with 0.5 mg/kgand 2 mg/kg of Compound YH-52 per day, and the mice in the control groupwere given the same volume of normal saline containing DMSO. The bodyweight of the mice and the length and width of the tumors were measuredand recorded every 3 days. The subcutaneous tumors were taken and thenphotographed at the end of the experiment. The tumor volumes werecalculated according to the equation: volume=length×width²×π/6, and thencounted. The experimental results are shown in FIG. 2A, FIG. 2B and FIG.2C.

Experiment of Intraperitoneal Injection Administration in Combinationwith Enzalutamide:

When the tumor grew to about 100 mm³, the mice were randomly dividedinto 4 groups to make sure that the average tumor volume was the same.The mice in the dosing group were intraperitoneally injected with 0.1mg/kg of Compound YH-52 per day, the mice in the positive drug groupwere intraperitoneally injected with 10 mg/kg of enzalutamide per day,the mice in the combination group were intraperitoneally injected with0.1 mg/kg of Compound YH-52 and 10 mg/kg of enzalutamide per day, andthe mice in the control group were given the same volume of normalsaline containing DMSO. The body weight of the mice and the length andwidth of the tumor were measured and recorded every 3 days. Thesubcutaneous tumors were taken and then photographed at the end of theexperiment. The tumor volumes were calculated according to the equation:volume=length×width²×π/6, and then counted. The experimental results areshown in FIG. 3A, FIG. 3B, FIG. 3C and FIG. 3D.

(3) Experimental Results

As shown in FIG. 2A, FIG. 2B and FIG. 2C, Compound YH-52 couldsignificantly inhibit tumor growth in mice at an oral gavage dose of 0.5mg/kg and 2 mg/kg. As shown in FIG. 3A, FIG. 3B, FIG. 3C and FIG. 3D,compared to the positive drug enzalutamide (ENZ), Compound YH-52administrated by intraperitoneal injection (at a dose of 0.1 mg/kg)showed a greater efficacy, and has a significant synergistic effect at adose of 0.1 mg/kg when being administrated by intraperitoneal injectionin combination with ENZ (10 mg/kg).

It is foreseeable that pharmaceutically acceptable salts of the compoundcan produce the same or similar activity.

1. A daphnane diterpenoid compound of formula I or formula II:

wherein: in the formula I and the formula II, a bond between C-1 and C-2is a single or double bond, a bond between C-6 and C-7 is a single ordouble bond, and a bond between C-15 and C-16 is a single or doublebond; in the formula I and the formula II, R₁ is hydrogen or hydroxyl orR₁ is absent when a double bond is formed between C-1 and C-2; in theformula I and the formula II, R₂ is hydrogen, hydroxyl, carbonyl,benzoyl, or acetyl; in the formula I and the formula II, R₃ is hydrogen,hydroxyl, acetyl, isovaleryl, crotonyl, or benzoyl; in the formula I andthe formula II, R₄ is hydrogen, hydroxyl, acetyl, isobutyryl,2-thienylcarbonyl, benzoyl, or palmitoyl; in the formula I and theformula II, R₅ is hydroxyl, fluorine, chlorine, bromine, or iodine, orforms a ternary epoxy with R₆ or R₆ is absent when a double bond isformed between C-6 and C-7; in the formula I and the formula II, R₆ ishydrogen, hydroxyl, fluorine, chlorine, bromine, or iodine, or forms aternary epoxy with R₅ or R₅ is absent when a double bond is formedbetween C-6 and C-7; in the formula I, R₇ is methyl, phenyl, nonanyl,(1E, 3E)-nonadienyl, (1E, 3Z)-nonadienyl, or (1E, 3E, 5E)-nonatrienyl;in the formula II, R₇ is hydrogen, benzoyl, acetyl, decanoyl, (2E,4E)-decadienoyl, (2E, 4Z)-decadienoyl, or (2E, 4E, 6E)-decatrienoyl; inthe formula I and the formula II, R₈ is hydrogen or hydroxyl or R₈ isabsent when a double bond is formed between C-15 and C-16; and in theformula I and the formula II, R₉ is hydrogen, hydroxyl, acetyl, benzoyl,isobutyryl, butyryl, or propionyl.
 2. The daphnane diterpenoid compoundaccording to claim 1, wherein the daphnane diterpenoid compound isselected from the group consisting of: a compound of formula (III):

wherein the compound of formula (III) is selected from the groupconsisting of: Compound R₁ R₂ R₃ R₄ R₇ R₈ R₉ YH-1 Δ1 ═O OH OH Ph Δ16 OBuYH-2 Δ1 ═O OH OH Ph Δ16 OiBu YH-3 Δ1 ═O OH OH Ph Δ16 OProp YH-4 Δ1 ═O OHOH Ph Δ16 OH YH-5 Δ1 ═O OH OG Ph Δ16 OAc YH-6 Δ1 ═O OH OH Ph Δ16 OAcYH-7 Δ1 ═O OAc OAc Ph Δ16 OAc YH-8 Δ1 ═O OH OAc Ph Δ16 OAc YH-9 Δ1 ═O OHOBz Ph Δ16 OAc YH-10 Δ1 ═O OH OS Ph Δ16 OAc YH-11 Δ1 ═O OH OH Ph Δ16 OBzYH-12 Δ1 ═O OAc OAc Ph Δ16 OBz YH-13 Δ1 ═O OH OAc Ph Δ16 OBz YH-14 Δ1 ═OOH OH A Δ16 OH YH-15 Δ1 ═O OH OH A Δ16 OBu YH-16 Δ1 ═O OH OH A Δ16 OBzYH-17 Δ1 ═O OH OH D Δ16 OBz YH-18 Δ1 ═O OAc OAc A Δ16 OBz YH-19 Δ1 ═O OHOAc A Δ16 OBz YH-20 Δ1 ═O OH OBz A Δ16 OBz YH-21 Δ1 ═O OH OS A Δ16 OAcYH-22 Δ1 ═O OH OH A Δ16 OAc YH-23 Δ1 ═O OAc OAc A Δ16 OAc YH-24 Δ1 ═O OHOAc A Δ16 OAc YH-25 Δ1 ═O OH OBz A Δ16 OAc YH-26 Δ1 ═O OH OS A Δ16 OAcYH-27 Δ1 ═O OH OG A Δ16 OAc YH-28 Δ1 ═O OH OG D Δ16 OBz YH-29 Δ1 ═O OHOH B Δ16 OBz YH-30 Δ1 ═O OH OH Me Δ16 OAc YH-31 Δ1 ═O OH OH B Δ16 OAcYH-32 Δ1 ═O OH OH D Δ16 OAc YH-33 Δ1 ═O OH OH F Δ16 OAc YH-34 Δ1 ═O OHOH F Δ16 OBz YH-35 αH ═O OH OH Ph Δ16 OAc YH-36 αH ═O OH OH Ph Δ16 OBzYH-37 Δ1 ═O OH OH Da H OAc YH-38 αH ═O OH OH Da H OAc YH-39 αH ═O OH OHPh H OBz YH-40 Δ1 ═O OH OH Ph H OBz YH-41 Δ1 ═O OH OH Ph Δ16 H YH-42 αHβO(CO)E OH OH Ph Δ16 H YH-43 αH βOH OH POE Ph Δ16 H;

a compound of formula (IV):

wherein the compound of formula (IV) is selected from the groupconsisting of: Compound R₃ R₄ R₅ R₆ R₇ R₉ YH-44 H OH Δ6 H Me OBz YH-45OH OBz βCl αOH Ph OAc YH-46 OH OBz αOH βCl Ph OAc YH-47 OH OH βCl αOH PhOAc YH-48 OH OH αOH βCl Ph OAc YH-49 OH OH βCl αOH Ph OBz YH-50 OH OHαOH βCl Ph OBz YH-51 OH OH αOH αOH Ph OBz YH-52 OH OH βBr αOH Ph OBzYH-53 OH OH αOH βBr Ph OBz;

a compound of formula (V):

wherein the compound of formula (V) is selected from the groupconsisting of: Compound R₇ R₉ YH-54 COA OAc YH-55 COA OBz YH-56 Bz OBz;

 and a compound of formula (VI):

wherein the compound of formula (VI) is selected from the groupconsisting of: Compound R₇ R₉ YH-57 H OBz YH-58 H OAc YH-59 COD OAcYH-60 Bz OBz YH-61 COA OH YH-62 COA OAc YH-63 COA OBz YH-64 COA H;

 and wherein in formula (III), formula (IV), formula (V), and formula(VI):

Δ1 is when R₁ is absent and a double bond is formed between C-1 and C-2;Δ6 is when R₅ is absent and a double bond is formed between C-6 and C-7;Δ16 is when R₈ is absent and a double bond is formed between C-15 andC-16; and α is when the identified substituent is on the opposite faceof the hydroxyl group of the contiguous ring; and β is when theidentified substituent is on the same face as the hydroxyl group of thecontiguous ring.
 3. The daphnane diterpenoid compound according to claim2, wherein the daphnane diterpenoid compound is selected from the groupconsisting of YH-6, YH-11, YH-16, YH-17, YH-22, YH-35, YH-36, YH-47,YH-48, YH-49, YH-50, YH-52, and YH-53.
 4. The daphnane diterpenoidcompound according to claim 1, wherein the daphnane diterpenoid compoundis a pharmaceutically acceptable derivative thereof.
 5. The daphnanediterpenoid compound according to claim 4, wherein the pharmaceuticallyacceptable derivative is a salt thereof.
 6. A composition comprising atleast one daphnane diterpenoid compound according to claim 1 and/or apharmaceutically acceptable derivative thereof.
 7. A compositioncomprising at least one daphnane diterpenoid compound selected from thegroup consisting of YH-8, YH-9, YH-10, YH-19, YH-20, YH-21, YH-24,YH-25, YH-26, YH-30, YH-33, YH-34, YH-37, YH-38, YH-39, YH-45, YH-46,YH-47, YH-48, YH-49, YH-50, YH-52, YH-53, YH-56, YH-57, YH-60, and YH-61according to claim 2 and/or a pharmaceutically acceptable derivativethereof.
 8. A composition for treatment or adjuvant treatment of acastration-resistant prostate cancer, comprising at least one daphnanediterpenoid compound according to claim 1, or a pharmaceuticallyacceptable derivative thereof.
 9. A composition for treatment oradjuvant treatment of a castration-resistant prostate cancer, comprisingat least one daphnane diterpenoid compound selected from the groupconsisting of YH-6, YH-11, YH-16, YH-17, YH-22, YH-35, YH-36, YH-8,YH-9, YH-10, YH-19, YH-20, YH-21, YH-24, YH-25, YH-26, YH-30, YH-33,YH-34, YH-37, YH-38, YH-39, YH-45, YH-46, YH-47, YH-48, YH-49, YH-50,YH-52, YH-53, YH-56, YH-57, YH-60, and YH-61 according to claim 2, or apharmaceutically acceptable derivative thereof.
 10. The compositionaccording to claim 9, further comprising at least one compound having atherapeutic effect on a castration-resistant prostate cancer.
 11. Amethod for treatment or adjuvant treatment of a castration-resistantprostate cancer in a patient, comprising: determining whether thepatient suffers from the castration-resistant prostate cancer; andadministering to the patient a therapeutic amount of a daphnanediterpenoid compound according to claim 1 or a pharmaceuticallyacceptable salt, a solvate, or a co-crystal thereof.
 12. The methodaccording to claim 11, further comprising administering to the patientat least one compound having a therapeutic effect on thecastration-resistant prostate cancer.
 13. The method according to claim12, wherein the compound having the therapeutic effect on thecastration-resistant prostate cancer is selected from the groupconsisting of enzalutamide, abiraterone, cyclophosphamide, adriamycin,docetaxel, mitoxantrone, and combinations thereof.
 14. The compositionaccording to claim 8, wherein the composition further comprises at leastone compound having a therapeutic effect on a castration-resistantprostate cancer.
 15. The composition according to claim 14, wherein thecompound having the therapeutic effect on the castration-resistantprostate cancer is selected from the group consisting of enzalutamide,abiraterone, cyclophosphamide, adriamycin, docetaxel, mitoxantrone, andcombinations thereof.
 16. The composition according to claim 10, whereinthe compound having the therapeutic effect on the castration-resistantprostate cancer is selected from the group consisting of enzalutamide,abiraterone, cyclophosphamide, adriamycin, docetaxel, mitoxantrone, andcombinations thereof.
 17. A method for treatment or adjuvant treatmentof a castration-resistant prostate cancer in a patient, comprisingadministering to a patient in need thereof a therapeutic amount of thecomposition according to claim
 8. 18. The method according to claim 17,further comprising administering to the patient at least one compoundhaving a therapeutic effect on the castration-resistant prostate cancer.19. A method for treatment or adjuvant treatment of acastration-resistant prostate cancer in a patient, comprisingadministering to a patient in need thereof a therapeutic amount of thecomposition according to claim
 9. 20. The method according to claim 19,further comprising administering to the patient at least one compoundhaving a therapeutic effect on the castration-resistant prostate cancer.